| Oligosaccharide has the function of regulating intestinal flora,moistening the bowel and improving the immunity of human body.Therefore,it is often added to infant formula milk to help establish the intestinal flora of newborn babies.At present,galacto-oligosaccharide(GOS),fructo-oligosaccharide(FOS)and their combination are used most in infant formula milk.According to the National Standard GB 14880-14880,the total content of oligosaccharide composition must not exceed 6.45 g/100 g.Through market research,it could be found that the content of oligosaccharide of different infant formula milk brands are all within national standard,but there are wide differences between them.And at present,researches about how the oligosaccharides added quantity of the milk affects the intestinal probiotics are limited,so this is still a question that whether there will be prominent differences due to different oligosaccharides quantity added to infant formula milk,and on the basis of safety,how much oligosaccharides adding quantity can has the best effects on intestinal probiotics.So the test is intended to see the influences on intestinal probiotics according to different adding amounts of GOS and FOS in infant formula milk by testing the number of living bacterium,and following-up mouse tests so as to select the optimum adding amount,as the final oligosaccharide content in milk of this test.And the adding of prebiotics and the choice of consumers can have a theoretical basis according to this.This paper explored the influences on intestinal probiotics according to different adding amounts of oligosaccharides in infant formula milk by using the in vitro experiments which intended to settle the mixture ratio of oligosaccharide in milk,then test the bifidobacterium,lactobacillus acidophilus live bacteria and intestinal flora and moistening bowel function of the mice based on this mixture ratio.The research contents and results are as follows:(1)Determination of the mixture ratio of oligosaccharides in infant formula milk:By testing OD value,probiotics amount and pH value,the optimum matching ratio of 8:2 was selected for GOS and FOS.In this matching group,the amount of live bacteria of bifidobacterium and acidophilus could reach 4.18×10~9 cfu/ml and 6.63×10~9 cfu/ml,which is significantly higher than that of other proportions(P<0.05).So in subsequent trials,the mixture ratio of GOS and FOS was8:2.(2)Inflence of oligosaccharide amount in infant formula milk on the numbers of live bacteria of bifidobacterium and lactobacillus acidophilus:When there is different amount of oligosaccharide in infant formula milk,the numbers of living bacterium of bifidobacterium and lactobacillus acidophilus are also different.When the dosage of oligosaccharide is 6 g/100 g,the number of bacteria of bifidobacteria could reach 5.23×10~9 cfu/ml,and the number of live bacteria of lactobacillus acidophilus reaches 6.8×10~9 cfu/ml.When oligosaccharide amount is 5.5 g/100 g,the number of bifidobacterium is 4.93×10~9 cfu/ml,and the number of living bacterium of lactobacillus acidophilus is 8.6×10~9 cfu/ml.The number of live bifidobacterium and lactobacillus acidophilus bacteria in the two dose groups of are significantly higher than other groups(P<0.05),and there is no obvious difference between the two groups(P>0.05).In consideration of the spending,the 5.5 g/100 g oligosaccharide was selected as the high-dose group in the experiment.When oligosaccharides amount is 1 g/100 g,the number of living bacterium of bifidobacterium and lactobacillus acidophilus were 7.47×10~8 cfu/ml and 1.36×10~9 cfu/ml,which was significantly lower than other dose groups(P<0.05).Therefore,the 1 g/100 g oligosaccharides was seclectd as the low-dose group in the experiment.(3)Effect of oligosaccharide addition in infant formula on gut microbiota,blood routine parameters,and interferon-?(IFN-?)and interleukin-2(IL-2)levels in mice:when the amount of oligosaccharide in milk powder is 5.5 g/100 g,the number of living bacterium of bifidobacterium and lactobacillus can be significantly improved,reducing the number of living bacterium of enterobacter,enterococcus and gas capsule clostridium(P<0.05),which is a significant difference from other dose groups(P<0.05).When the dosage of oligosaccharide in milk is 3 g/100 g,the number of bifidobacteria and lactobacillus can be significantly increased,and the number of viable bacteria is significantly reduced(P<0.05).When the amount of oligosaccharide in milk powder was 1 g/100 g,there was no obvious effect on the number of live bacteria of bifidobacterium,lactobacillus,enterobacterium,enterococcus and clostridium(P>0.05).According to the test of routine blood index in mice and IFN-?and IL-2,the results are as follows:the formula containing low amount of oligosaccharide has no effect on the blood routine indexes in mice,and the high-dose group,compared with control group,can significantly improve the amount of IFN-?and IL-2 in mice serum(P<0.05).(4)Influence of oligosaccharide in infant formula milk on the purge function of mice:through the movement of intestine propulsion in mice,time of the first defecation and determination of defecation grain number and weight,the results are as follow:when milk contains 3 g/100 g and5.5 g/100 g of oligosaccharides,the rate of ink propulsion in mice,grain number and weight can be significantly increased.Time span of the first defecation can also be reduced(P<0.05),of which the laxative effect of 5.5 g/100 g oligosaccharidesis is significantly higher than that of 3 g/100 g.When the amount of oligosaccharide was 1 g/100 g,the increase of the ink rate,the number of excretion,the increase of weight and the shortened time of the initial defecation are not obviously effected.The above test results conclude that different oligosaccharides amount in infant formula milk have different effects on intestinal prebiotic function and that increased amount of oligosaccharides can obviously improve the intestinal prebiotic function.The oligosaccharides content in milk is 5.5 g/100 g,which is the optimum amount in the experiment and the ideal oligosaccharides amount in the tested milk.(5)Sensorial comparison of the infant formula milk in the final test with none-oligosaccharide milk produced by the same craft concludes that when the mix proportion of oligosaccharides added in the milk powder is 8:2 with 5.5 g/100 g oligosaccharides,there will be no negative effect on the smell,soakage,shape,and color of the milk. |
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